Mouse CRISPR Knockout Pooled Library (Gouda)
(Pooled Library #136987)

• Purpose:

  The mouse CRISPR Gouda lentiviral pooled library was designed for assays with limited cell numbers.

  This library has two guides per gene and is compatible with the Brie library (Addgene #73633).

• Vector Backbone:

  pRDA_118 (Plasmid #133459) - does not express Cas9

• Depositing Labs: John Doench
• Publication: DeWeirdt et al Nat Commun. 2020 Feb 6;11(1):752. doi: 10.1038/s41467-020-14620-6.

Library Details

• Species: Mouse
• Genes targeted: 21,601
• gRNAs: 44,155
• Controls: 500 non-targeting controls, 500 one-intergenic site targeting controls
• Lentiviral Generation: 3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (PDF, 90 KB)
  • Sequencing Protocol (PDF, 162 KB)
• Depositor Data:
  • Gouda gRNA sequences (TXT, 1.5 MB)
  • Gouda Read Counts (TXT, 1.1 MB)
  • Gouda sgRNA Abundance Graph (PDF, 84 KB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

NOTE: Lentiviral vectors can recombine during library amplification, resulting in a band of ~1 kb containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the PuroR gene and will not be selected during screening. If recombination is observed after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.